title: Green tea constituents (-)-epigallocatechin-3-gallate (EGCG) and gallic acid induce topoisomerase I- and topoisomerase II-DNA complexes in cells mediated by pyrogallol-induced hydrogen peroxide.
abstract: Green tea and its major active constituent, (-)-epigallocatechin-3-gallate (EGCG), are in clinical trials for the prevention and treatment of several diseases such as cancer. DNA topoisomerase (topo) poisons are commonly prescribed anticancer drugs that kill cancer cells by inducing topo-DNA complexes. Using purified topoisomerases, previous in vitro studies have shown that EGCG induces the formation of topo-DNA complexes. Because the activity of a drug on purified topoisomerases does not always represent the activity in a cell, we have used an immunofluorescence technique that allows the visualisation of topo I- and topo II-DNA complexes produced in individual cells to evaluate the activity of EGCG on both enzymes. High levels of topo I- and topo II-DNA complexes were observed in K562 leukaemia cells exposed to EGCG. Similar levels of topo I- and topo II-DNA complexes were visualised in cells treated with gallic acid (GA) (the acid part of the EGCG ester). Pyrogallol (PG) also induced topo-DNA complexes with both enzymes, therefore suggesting that the activity of EGCG and GA is mediated by their PG moieties. Catalase prevented both the cytotoxicity and the formation of topo I- and topo II-DNA complexes induced by EGCG, GA, PG and myricetin (a PG-containing flavonoid recently shown to induce topo I- and topo II-DNA complexes in cells), indicating that hydrogen peroxide mediates these activities. Hydrogen peroxide induced topo I- and topo II (α and β)-DNA complexes in a time- and dose-dependent manner. The formation of topo I- and topo II-DNA complexes in cells exposed to hydrogen peroxide correlated well with the induction of apoptosis, suggesting that the topo-DNA complexes induced at long exposure times by the compounds tested in our study may be apoptotic topo-DNA complexes. Finally, we report results suggesting that PG-containing drugs may selectively kill tumour cells by generating hydrogen peroxide.
authors.affiliation: Institute for Cell and Molecular Biosciences, The Medical School, Newcastle University, Newcastle-upon-Tyne, NE2 4HH, UK.
authors.forename: Miguel
authors.initials: M
authors.lastname: López-Lázaro
authors.forename: José Manuel
authors.initials: JM
authors.lastname: Calderón-Montaño
authors.forename: Estefanía
authors.initials: E
authors.lastname: Burgos-Morón
authors.forename: Caroline A
authors.initials: CA
authors.lastname: Austin
mesh_terms: D000818:Animals; D055162:Biocatalysis / Q000187:drug effects; D002374:Catalase / Q000378:metabolism; D002392:Catechin / Q000031:analogs & derivatives* / Q000737:chemistry / Q000494:pharmacology; D016923:Cell Death / Q000187:drug effects; D002460:Cell Line; D004247:DNA / Q000378:metabolism*; D004264:DNA Topoisomerases, Type I / Q000378:metabolism*; D004250:DNA Topoisomerases, Type II / Q000378:metabolism*; D005419:Flavonoids / Q000737:chemistry / Q000494:pharmacology; D005707:Gallic Acid / Q000737:chemistry / Q000494:pharmacology*; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology*; D051379:Mice; D008954:Models, Biological; D011748:Pyrogallol / Q000737:chemistry / Q000494:pharmacology*; D013662:Tea / Q000737:chemistry; D013997:Time Factors
chemical_list: D005419:Flavonoids; D013662:Tea; D011748:Pyrogallol; D005707:Gallic Acid; C040015:myricetin; D002392:Catechin; D004247:DNA; D006861:Hydrogen Peroxide; C045651:epigallocatechin gallate; D002374:Catalase; D004264:DNA Topoisomerases, Type I; D004250:DNA Topoisomerases, Type II